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. 2012 Aug 10;24(8):3420–3434. doi: 10.1105/tpc.112.099861

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© 2012 American Society of Plant Biologists. All rights reserved.

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Figure 8. — A2L Associates with BSL1 but Does Not Promote an Interaction between BSL1 and R2.

(A) Immunoblots showing A2L coimmunoprecipitates with tomato BSL1 in planta. FLAG:A2L was transiently expressed alone or with SlBSL1:GFP in N. benthamiana. Immunoprecipitates obtained with anti-FLAG or anti-GFP antiserum and total protein extracts were immunoblotted with appropriate antisera. IP, immunoprecipitate; Rubisco, ribulose-1,5-bisphosphate carboxylase/oxygenase.

(B) Y2H analysis demonstrating that A2L (excluding the signal peptide) and the C-terminal effector domain of A2L (amino acids 66 to 116) interact with potato BSL1 (St-BSL1) in vivo. Both LacZ (blue color) and His3 (providing growth on medium lacking His [-his]) reporter genes were activated. Empty vector (pDEST32) and effector PITG_08949 (Gilroy et al., 2011) acted as negative controls (no reporter activity).

(C) Immunoblots showing A2L does not promote interaction of Sl-BSL1 with R2. FLAG:A2L, SlBSL1:myc, and GFP:SdR2 were transiently expressed in combination in N. benthamiana. Immunoprecipitates obtained with anti-FLAG or anti-GFP antiserum and total protein extracts were immunoblotted with appropriate antisera. PS, Ponceau stain.